抗鸡IgG重链单克隆抗体的制备与应用  

Preparation and Application of Monoclonal Antibodies Against Heavy Chain of Chicken Serum IgG

作　　者：辛颜彬[1] 刘景兰[1] 孟玲珍[1] 魏云玲[1] 孙继国 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100850

出　　处：《中国兽医杂志》1996年第3期6-7,共2页Chinese Journal of Veterinary Medicine

摘　　要：以提纯鸡IgG做抗原免疫Balb/c小鼠,取鼠细胞在-PEG_1000作用下与小鼠骨髓瘤细胞(Sp_2/o-Ag14)融合,采用间接免疫荧光(IFA)和酶联免疫吸附试验(ELISA)检测上清抗体,阳性孔经有限稀释法进行细胞克隆。共获得7株分泌抗鸡IgG单克隆抗体的杂交瘤细胞(2H_8、4D_8、1B_7、2A_7、2B_3、1G_12、3D_12),将这些细胞分别接种同系小鼠制备出腹水抗体,ELISA效价可达10~5～10~6,其中4株IFA效价为1∶400～1∶800,另3株为阴性。Ig亚类鉴定结果表明:除1G_12为lgG_(2a)外,其余均为IgG_1。经阻断试验、琼脂扩散试验和免疫印迹试验(Western blot)证明了单克隆抗体的特异性。上述杂交瘤细胞经3～6个月稳定传代后冻存于液氮中,1年后复苏培养,其分泌抗体的功能未见改变。The monoclonal antibodies (McAbs) against heavy chain of IgG from chicken sera were prepared by hybridoma techniques. The McAbs were generated by fusing Sp2/O- Ag^(14) mouse myeloma cells and splenocells from BALB/C mice immunized with the antigen of purified heavy chain of chiken serum IgG in the presence of PEG_(1000). The antibodies in hybridorna culture supernatant were detected by indirect immunofluorescence assay(IFA) and enzyme-linked immunosorbent assay(ELISA), then the positive hybridoma were cloned by the technique of limiting dilution. Seven cell lines (2H_8, 4D_8, 1B_7, 2A_7, 2B_3, 1G_(12), 3D_(12))of hybridoma secreting McAbs against heavy chain of chicken serum IgG were established, and the ascites were produced. The dilution titers of tbe McAbs were 1:10~5～1:10~6 in ELISA method. For the titers of IFA, four of the seven McAbs were 1:400～1:800, and the other three were negtive. The experiment results indicated that the immunoglobulin subclass of 1C_(12) was IgG_(2a), and the other six McAbs were IgG_1. The sensitivity and specificity of the McAbs were demonstrated by westernblot, agar diffusion and blocking test. Hybridoma cells fairly reserved the antibody—producing capacity after stable passaage for three to six months, and stored in liquid nitrogen for more than one year.

关 键 词：单克隆抗体 鸡血清 免疫球蛋白G 制备 

分 类 号：S852.43[农业科学—基础兽医学]